Human Cell Lines-Cell Lines-InCellGene LLC.-CellLines,Biomedicine reasearch Product,Cell Biology,Cell Culture,293T Cell,Media

Organism	Homo sapiens, human
Tissue	bone
Cell Type	osteoblast; SV40 large T antigen transfected
Product Format	frozen
Culture Properties	adherent
Biosafety Level	2 [Cells contain SV40 viral sequences] Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Age	fetus
Applications	The cells provide a homogenous, rapidly proliferating model system for studying normal human osteoblast differentiation, osteoblast physiology, and hormonal, growth factor, and other cytokine effects on osteoblast function and differentiation.
Disclosure	This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.

Karyotype	diploid, 43%; tetraploid, 57%
Derivation	This line was established by transfection of limb tissue obtained from a spontaneous miscarriage with the temperature sensitive expression vector pUCSVtsA58 and the neomycin resistance expression vector pSV2-neo. Clones were selected in the presence of 0.6 mg/mL G418.
Antigen Expression	SV40 T antigen
Genes Expressed	alkaline phosphatase
Cellular Products	alkaline phosphatase
Comments	Cells grown at a permissive temperature of 33.5°C exhibit rapid cell division (Doubling time of 36 hrs), whereas little cell division occurs at a restrictive temperature of 39.5°C (Doubling time of 96 hrs). The cells have the ability to differentiate into mature osteoblasts expressing the normal osteoblast phenotype. At the restrictive temperatures, cell division is slowed, differentiation increases, and a more mature osteoblast phenotype is produced.

Complete Growth Medium	The base medium for this cell line is a 1:1 mixture of Ham''''s F12 Medium Dulbecco''''s Modified Eagle''''s Medium,with 2.5 mM L-glutamine (without phenol red). To make the complete growth medium, add the following components to the base medium: 0.3 mg/ml G418; fetal bovine serum to a final concentration of 10%.
Subculturing	Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 34°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 34°C. Subcultivation Ratio: A subcultivation ratio of 1:4 is recommended Medium Renewal: Every 2 to 3 days
Cryopreservation	culture medium, 72%; additional fetal bovine serum, 20%; DMSO, 8%
Culture Conditions	Temperature: 34°C